Accelerated elimination of N. brasiliensis from the small intestine after auto-anti-IgE induction.

Immunization of rats with a purified IgE myeloma (IR2) induced an auto-anti-IgE response. Such treatment inhibited total IgE levels in the serum of conventional IgE-producing rats (Marshall & Bell, 1985) and increased the number of mucosal mast cells (MMC) in the intestine. The present study has investigated the ability of auto-anti-IgE induction to influence the course of a Nippostrongylus brasiliensis infection, to modify IgE synthesis, or to affect the number of MMC in the intestine following infection. Auto-anti-IgE induction was found to have a surprising effect on worm elimination. IR2-immunized rats were able to rid themselves of this nematode with an accelerated tempo--a small but significant effect after primary infection, but a substantial enhancement of worm loss after reinfection. Auto-anti-IgE induction was not able to prevent the typical increase in IgE that accompanies an N. brasiliensis infection, nor did it alter the helminth-induced intestinal mastocytosis. When MMC degranulation was measured by assaying the serum levels of a specific rat mast protease (RMCP II) following secondary infection, the amount of RMCP II released was less in auto-anti-IgE-producing rats. These findings have implications for the importance of IgE, MMC and other cells of inflammation in an anti-parasitic response.     

Expression of luminal and basal cytokeratins in human breast carcinoma

We have examined basal and luminal cell cytokeratin expression in 1944 cases of invasive breast carcinoma, using tissue microarray (TMA) technology, to determine the frequency of expression of each cytokeratin subtype, their relationships and prognostic relevance, if any. Expression was determined by immunocytochemistry staining using antibodies to the luminal cytokeratins (CKs) 7/8, 18 and 19 and the basal markers CK 5/6 and CK 14. Additionally, assessment of alpha-smooth muscle actin (SMA) and oestrogen receptor status (ER) was performed. The vast majority of the cases showed positivity for CK 7/8, 18 and 19 indicating a differentiated glandular phenotype, a finding associated with good prognosis, ER positivity and older patient age. In contrast, basal marker expression was significantly related to poor prognosis, ER negativity and younger patient age. Multivariate analysis showed that CK 5/6 was an independent indicator for relapse free interval. We were able to subgroup the cases into four distinct phenotype categories (pure luminal, mixed luminal/basal, pure basal and null), which had significant differences in relation to the biological features and the clinical course of the disease. Tumours classified as expressing a basal phenotype (the combined luminal plus basal and the pure basal) were in a poor prognostic subgroup, typically ER negative in most cases. These findings provide further evidence that breast cancer has distinct differentiation subclasses that have both biological and clinical relevance.     

Phenotype delineation of ZNF462 related syndrome

Zinc finger protein 462 (ZNF462) is a relatively newly discovered vertebrate specific protein with known critical roles in embryonic development in animal models. Two case reports and a case series study have described the phenotype of 10 individuals with ZNF462 loss of function variants. Herein, we present 14 new individuals with loss of function variants to the previous studies to delineate the syndrome of loss of function in ZNF462. Collectively, these 24 individuals present with recurring phenotypes that define a multiple congenital anomaly syndrome. Most have some form of developmental delay (79%) and a minority has autism spectrum disorder (33%). Characteristic facial features include ptosis (83%), down slanting palpebral fissures (58%), exaggerated Cupid's bow/wide philtrum (54%), and arched eyebrows (50%). Metopic ridging or craniosynostosis was found in a third of study participants and feeding problems in half. Other phenotype characteristics include dysgenesis of the corpus callosum in 25% of individuals, hypotonia in half, and structural heart defects in 21%. Using facial analysis technology, a computer algorithm applying deep learning was able to accurately differentiate individuals with ZNF462 loss of function variants from individuals with Noonan syndrome and healthy controls. In summary, we describe a multiple congenital anomaly syndrome associated with haploinsufficiency of ZNF462 that has distinct clinical characteristics and facial features.     

Phenotypic analysis of peripheral blood gammadelta T lymphocytes and their targeting by human immunodeficiency virus type 1 in vivo

There is increasing evidence that a wider range of lymphoid cell types other than CD4(+) T helper lymphocytes are infected with HIV-1 in vivo, including CD8 lymphocytes, natural killer cells, and reticulodendritic cells. Each potentially contributes to the reservoir of infected cells that resist antiviral treatment and to the impairment of immune responses in AIDS. By quantitative PCR for HIV proviral sequences we have now obtained evidence for substantial infection of gammadelta lymphocytes, contributing 3-45% of the proviral load in peripheral blood. A large proportion of gammadelta lymphocytes constitutively expressed the chemokine receptors CCR5 and CXCR4, with evidence for marked up-regulation of CD8 in samples from HIV-infected individuals, corresponding to an activated phenotype. That gammadelta lymphocytes might be susceptible to HIV infection was investigated using in vitro infectivity assays of recombinant HIV-expressing green fluorescent protein, followed by flow cytometry. gammadelta, CD4, and CD8 lymphocytes were each productively infected, with gammadelta lymphocytes showing the greatest susceptibility. For each cell type, blocking assays with an anti-CD4 monoclonal antibody indicated that entry was CD4-dependent.     

Quantitative measurement of cortical surface features in localization-related temporal lobe epilepsy

Differences in cortical surface features between healthy controls (n = 48) and patients with temporal lobe epilepsy (n = 46), ages 14-59, were characterized by means of advanced quantitative MRI processing techniques. Cortical surface features of interest included gyral and sulcal curvature, cortical depth, and total cortical surface area. Epilepsy patients and controls differed on measures of gyrification; the abnormalities generalized despite the focal nature of the primary epileptic process. Changes in cortical surface features were associated with increasing chronological age in both groups. Abnormalities in gyrification were associated with cognitive performance and with other morphometric measurements (e.g., surface cerebral spinal fluid). These findings are related to the literature regarding morphometric changes associated with temporal lobe epilepsy and normal aging.     

Vitamin A Levels and Immunity in Humans

In animal studies, vitamin A deficiency induces a shift from type 2 (humoral) to type 1 (cellular) cytokines; there are no similar data for humans. Control of human immunodeficiency virus (HIV) and Mycobacterium tuberculosis infections requires type 1 cytokine (cellular) immunity. These infections and vitamin A deficiency are highly prevalent in Africa. We therefore examined the interactions among serum vitamin A levels, immune parameters, HIV infection status, Mycobacterium bovis BCG vaccine scarring (as an indicator of a type 1 cytokine profile), and clinical findings for 70 hospitalized children in Malawi, Africa. Directly conjugated monoclonal antibodies and flow cytometry were used to assess cell-specific cytokine production by peripheral blood monocytes and lymphocyte subpopulations. The statistical techniques employed included nonparametric statistics and logistic regression analyses. Thirty percent of the participants had severe vitamin A deficiency (<10 μg/dl), 34% had moderate deficiency (10 to <20 μg/dl), and 36% had normal levels (≥20 μg/dl). Vitamin A levels were lower for HIV-positive than for HIV-negative children (median, 10 and 17 μg/dl, respectively). Vitamin A-deficient children (<20 μg/dl) were more likely than non-vitamin A-deficient children to have higher proportions of natural killer (NK) cells (median, 8.3 and 5.2%, respectively) and lower ratios of interleukin-10-producing monocytes to tumor necrosis factor alpha-producing monocytes after induction (median, 1.0 and 2.3, respectively). Vitamin A-deficient children were also more likely than non-vitamin A-deficient children to exhibit respiratory symptoms (47% versus 12%) and visible BCG vaccine scars (83% versus 48%), which are indicative of a type 1 response to vaccination. Vitamin A status did not vary with gender, age, incidence of malaria parasitemia, blood culture positivity, or rates of mortality (6% of vitamin A-deficient children died versus 20% of non-vitamin A-deficient children). Lower vitamin A levels were associated with a relative type 1 cytokine dominance and proportionately more NK cells, both of which may be somewhat beneficial to persons who are exposed to HIV, M. tuberculosis, or other type 1 pathogens.     

Prion protein accumulation and neuroprotection in hypoxic brain damage

The function of the normal conformational isoform of prion protein, PrP(C), remains unclear although lines of research have suggested a role in the cellular response to oxidative stress. Here we investigate the expression of PrP(C) in hypoxic brain tissues to examine whether PrP(C) is in part regulated by neuronal stress. Cases of adult cerebral ischemia and perinatal hypoxic-ischemic injury in humans were compared with control tissues. PrP(C) immunoreactivity accumulates within neuronal processes in the penumbra of hypoxic damage in adult brain, and within neuronal soma in cases of perinatal hypoxic-ischemic injury, and in situ hybridization analysis suggests an up-regulation of PrP mRNA during hypoxia. Rodents also showed an accumulation of PrP(C) in neuronal soma within the penumbra of ischemic lesions. Furthermore, the infarct size in PrP-null mice was significantly greater than in the wild type, supporting the proposed role for PrP(C) in the neuroprotective adaptive cellular response to hypoxic injury.     

Creutzfeldt-Jakob disease (CJD) with a mutation at codon 148 of prion protein gene: relationship with sporadic CJD

Creutzfeldt-Jakob disease (CJD), the most common human prion disease, includes sporadic (s) and familial (f) forms. Regardless of etiology, both forms are thought to share the pathogenic mechanism whereby the cellular prion protein (PrP(C)) converts into its pathogenic isoform (PrP(Sc)). While PrP(C) conversion is thought to be random in sCJD, conversion in fCJD is facilitated by the congenital presence of mutated PrP. Differences in PrP genotype (PRNP) and in conversion circumstances lead to PrP(Sc) with distinct characteristics that elicit different disease phenotypes. Here, we describe a case of fCJD with a substitution of histidine (H) for arginine (R) at codon 148 (R148H) and heterozygosity of the methionine/valine (M/V) polymorphic codon 129, with the 129M allele coupled with the mutation. The disease phenotype and all major characteristics of PrP(Sc) of fCJD(R148H) were virtually indistinguishable from those of sCJDMV2, which has features different from those of any other sCJD. Therefore, despite the differences in etiology, PRNP, and conversion process, the two forms of PrP(Sc) had similar characteristics. Furthermore, comparison of fCJD(R148H) with a recently reported case carrying R148H and homozygosity at codon 129 suggests that codon 129 coupled with the mutation as well as that located on the normal allele can modify major phenotypic and PrP(Sc) features of fCJD(R148H).     

T-helper subset function in the gut of rats: differential stimulation of eosinophils, mucosal mast cells and antibody-forming cells by OX8- OX22- and OX8- OX22+ cells.

Thoracic duct lymphocytes (TDL) collected 3 days after infection of rats with Trichinella spiralis (TS) and adoptively transferred into normal, uninfected recipients, increased the numbers of both mucosal mast cells (MMC) and eosinophils (EOS) in the intestine. The CD4+ T-helper cell population was separated into two subsets (OX22+ and OX22-) using OX22 monoclonal antibody (mAb) and panning techniques. After adoptive transfer of these T-helper subsets i.v., rats were challenged with TS 24 hr later. The intestine of recipient rats was examined histologically at intervals from Day 3 to Day 21. On Day 9 after transfer, OX22+ T helpers induced a substantial mastocytosis [94 +/- 3, mean +/- SE/villus crypt unit (VCU)], whereas the OX22- T-helper subset increased resident EOS numbers (60 +/- 2/VCU) compared to the challenge control (18 +/- 1 MMC, 27 +/- 1 EOS/VCU). The time of peak eosinophilia was advanced by 3-6 days for recipients of OX22- cells and that of mast cells by 9-12 days for recipients of OX22+ cells. The recipients of OX22-, but not OX22+, cells also showed a large increase in the numbers of B cells in the spleen and mesenteric lymph node (MLN) secreting antibody against adult TS. Recipients of OX22- cells displayed an even increase in EOS throughout the villi, lamina propria (LP) and muscularis, whereas in OX22+ cell recipients mast cells were only present in the lower villus and the epithelium just above the crypt as well as the muscularis layer. Only the CD4+ OX22- cell subset conferred protection against TS in the intestine. We conclude that the OX22+ and OX22- T-helper cells exert distinctive effects in the intestine on MMC and EOS. Because protection was established in the presence of an OX22- T-helper-induced eosinophilia but without a concurrent mastocytosis, the results suggest that MMC are probably not involved in expulsion of TS to terminate the primary infection.